LOCUS       Exported                7945 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor bar, complete sequence.
ACCESSION   MG687279
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7945)
  AUTHORS   Collier R, Thomson JG, Thilmony R.
  TITLE     GAANTRY, a versatile and robust Agrobacterium-based gene stacking 
            system generates high quality transgenic plants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7945)
  AUTHORS   Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit,
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 7945)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7945
                     /organism="Cloning vector GAANTRY P donor bar"
                     /mol_type="other DNA"
                     /db_xref="taxon:2201077"
     regulatory      complement(1..253)
                     /regulatory_class="terminator"
                     /note="nos terminator"
     terminator      1..253
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(285..836)
                     /codon_start=1
                     /gene="bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=bar
                     /note="confers herbicide resistance (BASTA, 
                     phosphinothricin)"
                     /protein_id="AWK49549.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(285..836)
                     /gene="bar"
                     /label=bar
     regulatory      complement(887..1439)
                     /regulatory_class="promoter"
                     /note="nos promoter"
     promoter        complement(887..1066)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(1473..1489)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1589..1644
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    complement(1681..1705)
                     /label=Agrobacterium T-DNA right border
                     /note="Agrobacterium T-DNA right border"
     CDS             2037..2831
                     /codon_start=1
                     /gene="nptIII"
                     /product="aminoglycoside phosphotransferase"
                     /label=nptIII
                     /note="confers resistance to kanamycin"
                     /protein_id="AWK49550.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            2037..2831
                     /gene="nptIII"
                     /label=nptIII
     misc_feature    3023..3128
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(3151..3167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3175..3191
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(3199..3229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3244..3265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3498..3943
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3944..5365
                     /codon_start=1
                     /gene="sacB"
                     /product="levansucrase"
                     /label=sacB
                     /note="confers sensitivity to sucrose"
                     /protein_id="AWK49551.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            3944..5365
                     /gene="sacB"
                     /label=sacB
     rep_origin      complement(5463..6051)
                     /direction=LEFT
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     CDS             complement(6222..7082)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin"
                     /protein_id="AWK49552.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6222..7082)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(7083..7187)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7213..7668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7810..7826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7835..7899
                     /label=TP901 attP
                     /note="TP901 attP"